Transporter-dependent cytotoxicity of antiviral drugs in primary cultures of human proximal tubular cells. (1st July 2018)

• Record Type: Journal Article
• Authors: Lash, Lawrence H.; Lee, Caroline A.; Wilker, Clynn; Shah, Vishal
• Is Part Of: Toxicology. Volume 404/405(2018)
• Page Start: 10
• Page End: 24
• Publication Date: 2018-07-01
• Subjects: ARV antiretroviral -- CHO Chinese Hamster Ovary -- CID collision-induced dissociation -- DMEM/F12 Dulbecco's Modified Eagle's Medium: Ham's F12 Medium -- DMF dimethylformamide -- DMSO dimethylsulfoxide -- GGT γ-glutamyltransferase -- HIV human immunodeficiency -- hPT human proximal tubular -- IS internal standard -- JAB apical-to-basolateral flux -- JBA basolateral-to-apical flux -- KIM-1 Kidney Injury Molecule-1 -- LC–MS/MS liquid chromatography-mass spectrometry/mass-spectrometry -- LDH lactate dehydrogenase -- MATE1 multidrug and toxin extrusion protein 1 -- MRM multiple reaction monitoring -- MRP multidrug resistance-associated protein -- NAG N-acetyl-β-d-glucosaminidase -- OAT organic anion transporter -- OATP organic anion-transporting polypeptide -- OCT organic cation transporter -- PBS phosphate-buffered saline -- P-gp P-glycoprotein (also called MDR1, multidrug resistance protein 1) -- PT proximal tubular -- TAF tenofovir alafenamide -- TDF tenofovir disoproxil fumarate -- TISP turbo ionspray; Antiviral drugs -- Nephrotoxicity -- Human proximal tubular cells -- Biomarkers -- Membrane transport; Toxicology -- Periodicals; Chemicals -- Physiological effect -- Periodicals; 615.9005
• DOI: 10.1016/j.tox.2018.05.002
• Languages: English
• ISSNs: 0300-483X

Abstract

Abstract: The role of plasma membrane transporters in the nephrotoxicity of two antiretroviral drugs, cidofovir and tenofovir, was studied in primary cultures of human proximal tubular (hPT) cells. Cells were grown on Transwell filter inserts to maintain epithelial polarity and access to either the apical or basolateral plasma membrane. The function of relevant membrane transporters, organic anion transporter 1 and 3 (OAT1/3), P-glycoprotein (multidrug resistance protein-1; P-gp or MDR1), and organic cation transporter 2 (OCT2), was validated by measurements of apical-to-basolateral and basolateral-to-apical fluxes of furosemide, digoxin, and metformin, respectively. Acute cytotoxicity of cidofovir (0, 10, 50, 150, or 300 μM) in the absence or presence of 500 μM probenecid, tenofovir disoproxil fumarate (0, 20, 90, 180, or 360 μM) in the absence or presence of 500 μM probenecid, or cisplatin (0, 20, 90, 180, or 360 μM) as a positive control in the absence or presence of 500 μM cimetidine, was assessed after 4-h incubations by determinations of release of lactate dehydrogenase (LDH), γ-glutamyltransferase (GGT), N -acetyl-β-d -glucosaminidase (NAG), or Kidney Injury Molecule-1 (KIM-1). Cell death generally agreed with each of the four biomarkers, was generally greater when cidofovir or tenofovir was added to the upper compartment, and was markedly diminished in the presence of the appropriate transport inhibitor. Additionally, the extent of cytotoxicity caused by the two antiviral drugs was similar to that caused by cisplatin. The results demonstrate the importance of plasma membrane transport of antiviral drugs to elicit cytotoxicity in the hPT cell.